Acidovorax citrulli Seed Inoculum Load Affects Seedling Transmission and Spread of Bacterial Fruit Blotch of Watermelon Under Greenhouse Conditions

Abstract

Infested seed are typically the primary source of inoculum for bacterial fruit blotch (BFB) of cucurbits. An inoculum threshold of 1 infested seed per 10,000 seeds is widely used in seed health testing for Acidovorax citrulli. However, the influence of seed inoculum load on BFB seedling transmission has not been elucidated. In this study, watermelon seedlots (128 seeds/lot) containing one seed inoculated with A. citrulli at levels ranging from 1 × 10¹ to 1 × 10⁷ CFU were used to investigate the effect of seed inoculum load on seedling transmission and spatiotemporal spread of BFB under greenhouse conditions. The relationship between A. citrulli seed inoculum load and frequency of BFB seedling transmission followed a sigmoidal pattern (R² = 0.986, P = 0.0047). In all, 100 and 96.6% of seedlots containing one seed with 1 × 10⁷ and 1 × 10⁵ CFU of A. citrulli, respectively, transmitted the pathogen to seedlings; in contrast, the proportion of seedlots that yielded BFB-infected seedlings was lower for lots with one seed infested with 1 × 10³ (46.6%) and 1 × 10¹ (16.7%) CFU of A. citrulli. The relationship between A. citrulli seed inoculum load and frequency of pathogen detection in seedlots using immunomagnetic separation combined with a real-time polymerase chain reaction assay also followed a sigmoidal pattern (R² = 0.997, P = 0.0034). Whereas 100% of samples from seedlots (10,000 seeds/lot) with one seed containing ≥1 × 10⁵ CFU tested positive for A. citrulli, 75% of samples from lots with one seed containing 1 × 10³ CFU tested positive for the pathogen, and only 16.7% of samples with one seed containing 10 CFU tested positive. Because disease transmission was observed for lots with just one seed containing 10 A. citrulli CFU, zero tolerance for seedborne A. citrulli is recommended for effective BFB management. The seedling transmission experiments also revealed that temporal spread of BFB in 128-cell seedling trays increased linearly with A. citrulli inoculum load (r² = 0.976, P = 0.0037). Additionally, the frequency of spatial spread of BFB from an inoculated seedling in the center of a planting tray to adjacent healthy seedlings over one-, two-, or three-cell distances was greater for lots with one seed infested with at least 1 × 10⁵ CFU than for lots with one seed infested at lower inoculum loads (1 × 10¹ and 1 × 10³ CFU/seed).